OBJECTIVE: Sexual prejudice and masculine gender role stress were examined as mediators of the associations between adherence to different male gender norms and aggression toward sexual minorities. This study also sought to extend past research to a community sample and employ multiple methods to assess aggression. METHOD: Participants were 199 heterosexual men between the ages of 18-30 who were recruited from a large southeastern United States city. Participants completed measures of adherence to male gender role norms, sexual prejudice, masculine gender role stress, and aggression toward sexual minorities. RESULTS: Associations between adherence to the status and antifemininity norms and aggression toward sexual minorities were mediated by sexual prejudice, but not masculine gender role stress. The portion of unique association between adherence to the antifemininity norm and aggression toward sexual minorities was about three times larger than the portion mediated by sexual prejudice and masculine gender role stress. CONCLUSIONS: Findings provide the first multivariate evidence from a community-based sample for determinants of aggression toward sexual minorities motivated by gender role enforcement. These data support intervention programming and preventative intervention studies aimed at reducing sexual prejudice and facilitating less stereotypic attitudes about the male gender role, particularly surrounding the antifemininity norm.